高粱根抗凝药效物质基础及作用机制研究

With the rapid development of Guizhou liquor industry centered on Moutai liquor, The planting area of Sorghum used for wine making has expanded rapidly in Guizhou, and Sorghum root resources are abundant. However, most of the agricultural production as waste incineration has not been effectively utilized. Sorghum root is also a traditional traditional Chinese medicine, which has been recorded in traditional Chinese medicine. However, its chemical composition and pharmacological research are not deep enough, which hinders its development as a resource of traditional Chinese medicine. In recent years, the applicant has been concerned about the discovery and utilization of new resources of Chinese medicine. In the previous study of the project group, it was found that the extract of sorghum root has significant anticoagulant effects, suggesting that it has good potential in the development of antithrombotic drugs. The anticoagulant activity of sorghum root material basis and mechanism are still not clear, this project is on the basis of preliminary studies using HPLC-QTOF /MS technology combined with the active tracking method, a systematic research on the sorghum root anticoagulant efficacy material basis, effect of the active substance on platelet aggregation and blood coagulation factors, evaluation system of sorghum root as the possibility of anti thrombosis drug development. The successful implementation of this project will lay a theoretical foundation for the high value utilization of sorghum root resources, and play a positive role in promoting regional economic development.

随着以茅台酒为核心的贵州白酒产业的快速发展，贵州省酒用高粱种植面积迅速扩大，高粱根资源十分丰富。但农业生产中多将其作为废弃物焚烧，未能得到有效利用。高粱根其实也是一味传统中药材，中医药典籍中均有记载。但由于化学成分及药理研究不深入，阻碍了其作为中药资源的深度开发。申请人近年来一直关注中药新资源的发现和利用，项目组前期研究发现，高粱根提取物具有显著的抗凝血作用，提示其在作为抗血栓药物开发方面具有较好的潜力。由于高粱根抗凝药效物质基础和作用机制仍不明确，本项目拟在前期研究基础上，采用HPLC-QTOF/MS技术结合活性追踪方法，对高粱根抗凝药效物质基础进行系统研究，探讨药效物质对血小板聚集及凝血因子影响的作用机制，系统评估高粱根作为抗血栓药物开发的可能性。本项目的顺利实施，将为高粱根资源高值化利用奠定理论基础，对推动地区经济发展起到积极的作用。

贵州省酒用高粱的大量种植，导致高粱根资源十分丰富。但农业生产中多将其作为废弃物焚烧，未能得到有效利用。高粱根其实也是一味传统中药材，中医药典籍中均有记载。但由于化学成分、药理及作用机制研究不深入，阻碍了其作为中药资源的深度开发。本项目在前期研究基础上，对有抗凝活性的高粱根醇提正丁醇萃取部位（BES）和水提醇沉上清部位（WEAE）进行再分段和抗血小板聚集活性筛选，得到活性组分群WEAE-M30%。UPLC-MS/MS分析结果显示，WEAE-M30%中化学成分主要有黄酮类、酚酸类、脂质类、有机酸类、生物碱类、木脂素及香豆素类等。从高粱根提取物中分离鉴定了40个单体化合物，其中有3个新化合物，22个首次从高粱属中分离得到，6个首次从高粱根中分离获得。筛选了单体化合物抗血小板聚集活性，得到多个活性化合物。探讨了1个活性组分和3个活性化合物抗血小板聚集的作用机制。WEAE-M30%对Collagen介导的血小板GPVI信号通路蛋白Src、PI3K通路蛋白AKT， MAPK信号通路蛋白p38、ERK蛋白的磷酸化水平有显著抑制作用，且对血小板内颗粒释放物ATP、P-selection、Ca2+的水平有显著抑制作用。Sorgholide A能显著抑制Collagen介导的血小板GPVI信号通路PLCγ2、Src，PI3K信号通路蛋白AKT，MAPK信号通路蛋白p38的磷酸化水平，显著抑制ATP、Ca2+的释放，促进cAMP、cGMP的释放；同时也显著抑制Thrombin介导的血小板GPVI信号通路Src，PI3K信号通路蛋白AKT，MAPK信号通路蛋白p38、ERK蛋白的磷酸化水平，显著降低ATP、Ca2+、CD62P、TXB2/6-keto-PGF1α的水平，促进cAMP的释放。紫杉氰糖苷能显著抑制GPVI信号通路SYK、Src，PI3K信号通路蛋白AKT，MAPK信号通路蛋白ERK的磷酸化水平，显著抑制ATP、TXB2/6-ketoPGF1α、Ca2+水平。琥珀酸能显著抑制GPVI信号通路SYK、Src，PI3K信号通路蛋白AKT，MAPK信号通路蛋白ERK、P38的磷酸化水平，且显著抑制ATP、Ca2+、CD62P、TXB2/6- ketoPGF1α水平，促进cAMP、cGMP的释放作用。项目的顺利实施，为高粱根资源高值化利用奠定了理论基础，对推动地区经济发展起到了积极作用。